Crystallized structure of type IV collagen NC1 domain hexamer

Abstract

The present invention provides a crystallized NC1 domain hexamer of Type IV collagen, and methods for making the crystal, wherein the NC1 domain hexamer is crystallized such that the three dimensional structure of the crystallized NC1 domain hexamer can be determined to a resolution of at least 3 Å or better. The present invention also provides a method for designing compounds to inhibit angiogenesis, tumor growth, tumor metastasis, endothelial cell adhesion and/or proliferation, and/or basal lamina assembly, comprising analyzing the three dimensional structure of a crystallized Type IV collagen NC1 domain hexamer produced by the methods of the invention, and identifying and synthesizing compounds that target regions of the NC1 domain that have been identified by the analysis as being important for type IV collagen heterotrimer and hexamer assembly. The present invention also provides novel polypeptides designed by the rational drug design methods of the present invention, based on an analysis of the type TV collagen NC1 hexamer structure disclosed herein.

Description

CROSS REFERENCE[0001]This application claims priority to U.S. Provisional Patent Application Ser. Nos. 60 / 308,523 filed Jul. 27, 2001; 60 / 351,289 filed Oct. 29, 2001; 60 / 366,854 filed Mar. 22, 2002; and 60 / 385,362 filed Jun. 3, 2002.STATEMENT OF GOVERNMENT INTEREST[0002]This work was supported by Grants DK18381 and DK53763 from the National Institutes of Health, and thus the U.S. government may have certain rights in the invention.FIELD OF THE INVENTION[0003]The present invention relates to the fields of crystallography, molecular biology, protein chemistry, angiogenesis, tumor growth and metastasis, and basement membrane assemblyBACKGROUND OF THE INVENTION[0004]The basement membrane (basal lamina) is a sheet-like extracellular matrix (ECM), which is a basic component of all tissues. The basal lamina provides for the compartmentalization of tissues, and acts as a filter for substances traveling between tissue compartments. Typically the basal lamina is found closely associated with ...

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Benefits of technology

[0014]In another aspect, the present invention provides novel polypeptides designed by the rational drug design methods of the present invention, based on an analysis of the type IV collagen NC1 hexamer structure disclosed herein. As a result of the information available from the crystal structure, it is possible to predict individual NC1 domain sequences that are critical for assembly of the type IV collagen heterotrimer and / or hexamer. Thus, it is also possible to design therapeutic polypeptides that will interfere with those interactions, and to inhibit assembly of the type IV collagen heterotrimer and / or the type IV collagen hexamer. Such therapeutic polypeptides can be used to inhibit or disrupt type IV collagen assembly, and thus are useful to inhibit angiogenesis, angiogenesis-mediated disorders, tumor growth, tumor metastasis, endothelial cell adhesion and / or proliferation, and basal lamina assembly.

Problems solved by technology

While this approach has led to some success, the number of growth factors known to play a role an angiogenesis is large.

Therefore, the possibility exists that growth factor antagonists may have only limited use in treating cancer, since tumors and associated inflammatory cells likely produce a wide variety of factors that can induce angiogenesis.

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